[ABSTRACT]AIM:ToinvestigatetheeffectofD4F,anapolipoproteinA-Imimeticpeptide,onoxidizedlow-density lipoprotein ( ox-LDL)-induced macrophage apoptosis and activation of caspase-12, a key molecule in endoplasmic reticulum stress ( ERS )-associated apoptotic pathway, and to elucidate the underlying molecular mechanisms. METHODS:RAW264.7 macrophages were pretreated with D4F (12.5, 25 and 50 mg/L), 4-phenylbutyric acid (PBA, 5 mmol/L) or diphenyleneiodonium ( DPI, 5 μmol/L) for 1 h and then treated with ox-LDL (100 mg/L) or tunicamycin ( TM, 4 mg/L) for 24 h.The cell viability and apoptosis were determined by MTT assay and TUNEL detection, respective-ly.The levels of malondialdehyde ( MDA) and reactive oxygen species ( ROS) in the cells and the activities of superoxide dismutase ( SOD) and nicotinamide adenine dinucleotide phosphate ( NADPH) oxidase were determined.The protein level of caspase-12 was examined by Western blot analysis.RESULTS: Similar to the ERS inhibitor PBA, D4F protected RAW264.7 macrophages from ox-LDL or TM ( an ERS inducer)-induced decrease in the viability and increase in apoptotic rate in a dose-dependent manner.Like DPI (an oxidative stress inhibitor), D4F significantly inhibited ox-LDL-induced ox-idative stress, as expressed by the decreased generation of ROS and MDA ( P<0.01) , the increased activity of SOD and the decreased activity of NADPH oxidase (P<0.05).Moreover, similar to PBA and DPI, D4F significantly suppressed ox-LDL-induced activation of caspase-12 in a concentration-dependent manner ( P<0.05) .Furthermore, D4F also inhibi-ted the caspase-12 activation induced by TM (P<0.05).CONCLUSION: D4F inhibits macrophage apoptosis induced by ox-LDL, and the mechanism is at least partially by reducing oxidative stress and inhibiting the activation of caspase-12.

BACKGROUNDHyperglycemia in brain and spinal cord could aggravate neurologic impairment. Recent studies showed that L-lysine monohydrochloride (LMH) could increase the insulin secretion and regulate the blood glucose level. The aim of the present study was to investigate the effects of LMH on pancreatic islet B cells, the levels of endogenous insulin and blood glucose in spinal cord injured rats.

METHODSForty male Wistar rats were divided into four groups, namely, normal control group, model group, high-dose LMH group (621.5 mg/kg equal to LMH 1/8 LD50), and low-dose LMH group (310.8 mg/kg equal to LMH 1/16 LD50). The model of spinal cord injured rat was established by hemi-transection at the lower right thoracic spinal cord. LMH was administered via intraperitoneal injection once spinal cord injury was produced in rats. All rats were sacrificed 48 hours after spinal cord injured. The effects of LMH on pancreatic islet B cells, the content of endogenous insulin, and the level of blood glucose were observed with immunohistochemical method, radioimmunoassay method, and biochemical analyzer, respectively.

RESULTSThe insulin immunohistochemical intensities of islet B cells were significantly weaker in model group than those in normal control group (P < 0.01). The levels of endogenous insulin were significantly lower and the blood glucose levels were significantly higher in model group than those in normal control group (P < 0.01). The insulin immunohistochemical intensities of islet B cells were significantly stronger in high-dose LMH group than those in model group (P < 0.05). In addition, we found that the levels of endogenous insulin were significantly higher and the blood glucose levels were significantly lower in high-dose LMH group than those in model group (P < 0.05). There were no significant differences in the insulin immunohistochemical intensities of islet B cells, the levels of endogenous insulin and the blood glucose between low-dose LMH group and model group (P > 0.05).

CONCLUSIONLMH, but dose-dependent, might participate in the regulation of pancreatic islet B cells, and then reduce the blood glucose levels in the spinal cord injured rats.

Animals ; Blood Glucose ; analysis ; Hyperglycemia ; etiology ; Insulin ; blood ; Lysine ; pharmacology ; Male ; Neuroprotective Agents ; pharmacology ; Rats ; Rats, Wistar ; Spinal Cord Injuries ; blood ; complications

OBJECTIVETo study the effects of prenatal exposure to Di-(2-ethylhexyl)-phthalate (DEHP) on genome-wide epigenetic alterations in ovary of adult offspring rat.

METHODSPregnant Wistar rats were randomly treated with DEHP (1000 mg/kg) or con oil at 12 - 17 days upon pregnance. DNA methylation changes in the ovary for the adult offsprings which were 70 days old were detected by Rat DNA methylation promoter plus CpG island arrays CpG island chip. Gene ontology (GO) method was performed to analyze the function of genes which were significantly different between exposed group and control group. Gene Igfbp1 (insulin-like growth factor binding protein 1) and Itga3 (integrin alpha 3) were randomly selected and the methylation status were verified by bisulfite genomic sequencing (BSP).

RESULTSThe methylation status were significantly different between exposed and control group in 406 genes (71 genes as hypermethylation and 335 genes as hypomethylation) (P < 0.05). GO analysis revealed that molecular transducer activity, cell part, cell, cellular process, multicellular organismal process, response to stimulus, biological regulation, regulation of biological process, reproduction, reproductive process, and rhythmic process were involved. The sequencing results were consistent with the data obtained by chips.

CONCLUSIONThis study provides evidence that prenatal exposure of DEHP may be associated with methylation changes on the genes in the rat ovary. Genes related to reproductive process have highly significant methylation changes, which may shed new light on mechanisms of reproductive and developmental toxicity after prenatal exposure to DEHP.

Animals ; CpG Islands ; genetics ; DNA Methylation ; Diethylhexyl Phthalate ; toxicity ; Female ; Genome ; Maternal Exposure ; Ovary ; pathology ; Pregnancy ; Prenatal Exposure Delayed Effects ; Rats ; Rats, Wistar

Objective To study long term renal function of Donation after citizen's deceased transplantation.Methods We compared the data of 38 subjects who got Delayed Graft Function(DGF) with 80 Immediate Graft Function (IGF) subjects underwent DCD transplantation in our hospital before June 2016.Evaluated the renal function by detecting the serum creatinine (sCr),the estimating glomerular filtration rate (eGFR) calculated with MDRD formula and urine protein at the 1,2,3 year post transplantation.Results Analyzed the serum eGFR of two groups,there was no significant differences at 1 and 2 year post transplantation,sCr of two groups showed no significant differences at 3 year (P =0.053)post transplantation,eGFR of two groups showed significant differences at 3 year (P =0.042)post transplantation and positive incidence of urine protein showed significant differences at 2 year (P =0.028)and 3 year (P =0.037)post transplantation.Conclusion DGFoccuring after DCD transplantation had an effect on long term renal function,.mainly on reducing of eGFR and increasing of urine protein positive rate 2 or 3 years after transplant.

OBJECTIVETo evaluate the association of major histocompatibility complex class I chain related gene A (MICA) antibodies with acute rejection (AR), chronic rejection (CR) and renal function after renal transplantation.

METHODSSerum MICA antibodies were detected with ELISA before and after transplantation with also examinations of panel reactive antibodies (PRA), serum creatinine, urine, graft ultrasound, lymphocyte subsets and the pathology of graft biopsy. The study was carried out in two parts to monitor MICA antibodies in acute and chronic rejections after renal transplantation.

RESULTSIn the first part of the study 18 of the 41 recipients experienced episodes of acute rejection, and the incidence rate was markedly higher in MICA(+) group than in MICA(-) group (P<0.05). Compared with the recipients with stable renal functions, the patients with acute graft rejection showed a significantly higher positivity rate of MICA antibodies. Postoperative MICA antibody monitoring showed that MICA antibody level increased gradually 2-3 days after the occurrence of acute rejection; anti-rejection treatment lowered serum creatinine to a normal level but MICA antibodies remained positive. In the second part, 21 of 40 patients had chronic graft rejection and showed significantly higher positivity rate of MICA than the patients with stable renal functions (P<0.05). In patients with chronic rejections, the serum creatinine levels were significantly higher in MICA(+) than in MICA(-) cases (P<0.05). Graft biopsy of all MICA(+) cases showed C4d deposition.

CONCLUSIONThe status of MICA antibodies can predict the occurrence and treatment outcomes of acute rejection, and also as one of the major causes of chronic graft rejection, they affect the long-term survival of the renal grafts.

Adolescent ; Adult ; Antibodies ; blood ; immunology ; Complement C4b ; metabolism ; Creatinine ; blood ; Follow-Up Studies ; Graft Rejection ; blood ; immunology ; pathology ; HLA Antigens ; immunology ; Histocompatibility Antigens Class I ; immunology ; Humans ; Kidney ; metabolism ; physiopathology ; Kidney Transplantation ; Peptide Fragments ; metabolism ; Young Adult

Postmortem interval (PMI) estimation is one of the most challenging problems in the field of forensic science. Vitreous humor is a hotspot which has been used for PMI estimation and postmortem chemical analysis in forensic pathology. In order to provide novel perspectives for the future research of PMI estimation using vitreous humor, the comparison between vitreous humor with other common body fluids, the effect of temperature on vitreous humor, vitreous humor detection method and data fitting method have been reviewed in this paper.

Objective To evaluate the association of major histocompatibility complex class I chain related gene A (MICA) antibodies with acute rejection (AR), chronic rejection (CR) and renal function after renal transplantation. Methods Serum MICA antibodies were detected with ELISA before and after transplantation with also examinations of panel reactive antibodies (PRA), serum creatinine, urine, graft ultrasound, lymphocyte subsets and the pathology of graft biopsy. The study was carried out in two parts to monitor MICA antibodies in acute and chronic rejections after renal transplantation. Results In the first part of the study 18 of the 41 recipients experienced episodes of acute rejection, and the incidence rate was markedly higher in MICA+group than in MICA-group (P<0.05). Compared with the recipients with stable renal functions, the patients with acute graft rejection showed a significantly higher positivity rate of MICA antibodies. Postoperative MICA antibody monitoring showed that MICA antibody level increased gradually 2-3 days after the occurrence of acute rejection; anti-rejection treatment lowered serum creatinine to a normal level but MICA antibodies remained positive. In the second part, 21 of 40 patients had chronic graft rejection and showed significantly higher positivity rate of MICA than the patients with stable renal functions (P<0.05). In patients with chronic rejections, the serum creatinine levels were significantly higher in MICA+than in MICA-cases (P<0.05). Graft biopsy of all MICA+cases showed C4d deposition. Conclusion The status of MICA antibodies can predict the occurrence and treatment outcomes of acute rejection, and also as one of the major causes of chronic graft rejection, they affect the long-term survival of the renal grafts.

Objective To evaluate the association of major histocompatibility complex class I chain related gene A (MICA) antibodies with acute rejection (AR), chronic rejection (CR) and renal function after renal transplantation. Methods Serum MICA antibodies were detected with ELISA before and after transplantation with also examinations of panel reactive antibodies (PRA), serum creatinine, urine, graft ultrasound, lymphocyte subsets and the pathology of graft biopsy. The study was carried out in two parts to monitor MICA antibodies in acute and chronic rejections after renal transplantation. Results In the first part of the study 18 of the 41 recipients experienced episodes of acute rejection, and the incidence rate was markedly higher in MICA+group than in MICA-group (P<0.05). Compared with the recipients with stable renal functions, the patients with acute graft rejection showed a significantly higher positivity rate of MICA antibodies. Postoperative MICA antibody monitoring showed that MICA antibody level increased gradually 2-3 days after the occurrence of acute rejection; anti-rejection treatment lowered serum creatinine to a normal level but MICA antibodies remained positive. In the second part, 21 of 40 patients had chronic graft rejection and showed significantly higher positivity rate of MICA than the patients with stable renal functions (P<0.05). In patients with chronic rejections, the serum creatinine levels were significantly higher in MICA+than in MICA-cases (P<0.05). Graft biopsy of all MICA+cases showed C4d deposition. Conclusion The status of MICA antibodies can predict the occurrence and treatment outcomes of acute rejection, and also as one of the major causes of chronic graft rejection, they affect the long-term survival of the renal grafts.

BACKGROUNDThe clinical significance of ischemic chest pain before acute ST-elevation myocardial infarction (STEMI) remains an interesting issue of investigation particularly in the era of percutaneous coronary intervention (PCI). This study aimed to assess the impact of angina prior to STEMI on short-term clinical outcomes in patients with acute STEMI undergoing primary PCI.

METHODSAmong a total of 875 consecutive patients with STEMI undergoing primary PCI, 292 had episodes of angina within 24 hours of STEMI (PA group) and the remaining 583 were free of anginal symptoms (non-PA group). Clinical characteristics, angiographic and procedural features, and in-hospital and 30-day outcomes were compared between the two groups.

RESULTSDiabetes was less common (17.5% vs. 23.3%, P = 0.04) and symptom-to-door time was shortened ((191.6 ± 96.8) minutes vs. (357.2 ± 341.9) minutes, P < 0.001) in the PA group than in the non-PA group. Patients with angina prior to STEMI had fewer totally or nearly totally occluded infarct-related artery (TIMI flow grade 0 - 1) at initial angiography (75.0% vs. 90.7%, P < 0.001), and achieved more TIMI flow grade 3 after primary PCI (84.2% vs. 78.2%, P = 0.04). These were associated with higher rates of overall procedural success (95.9% vs. 91.8%, P = 0.02) and of complete ST-segment resolution at 90 minutes after the procedure (51.7% vs. 40.3%, P = 0.001). During a 30-day clinical follow-up, the left ventricular ejection fraction was significantly improved ((53.0 ± 8.6)% vs. (51.1 ± 9.7)%, P = 0.002) and the primary endpoint of major adverse cardiac events was reduced in the PA group (7.2% vs. 12.7%, P = 0.01).

CONCLUSIONPresence of angina prior to acute STEMI is associated with better outcome at a 30-day clinical follow-up in patients undergoing primary PCI.

Acute Coronary Syndrome ; therapy ; Aged ; Angina, Unstable ; physiopathology ; Angioplasty, Balloon, Coronary ; Coronary Angiography ; Electrocardiography ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; therapy ; Prospective Studies

A gene replacement/disruption system of Amycolatopsis mediterranei U32 was developed based on the established electroporation conditions as well as appropriate selective markers. Through two-step selection, ahbas gene in U32 was replaced by a promoterless alpha-amylase gene constructed on the plasmid pDK110 of E. coli. The first single-crossover and the second double-crossover frequencies were approximately 0.5%-0.7% and 2%, respectively. Denaturation of the plasmid pDK110 increased the integration frequency about 7-10 folds, while electric shock treatment of the single-crossover recombinants increased the frequency of second crossover recombination about 5 folds. Employing denatured DNA fragments containing an apramycin-resistance gene flanked with regions of the respective genes, One-step disruption of rifO and amrA genes of U32 was also achieved with an efficiency of 30-50 transformants per microgram of DNA.
Actinomycetales ; drug effects ; genetics ; growth & development ; DNA, Bacterial ; genetics ; Drug Resistance, Microbial ; genetics ; Genes, Bacterial ; genetics ; Mutagenesis ; Nebramycin ; analogs & derivatives ; pharmacology ; Plasmids ; genetics ; Recombination, Genetic